Profiling humoral immune responses to P. falciparum infection with protein microarrays.
A complete description of the serological response following exposure of humans to complex pathogens is lacking and approaches suitable for accomplishing this are limited. Here we report, using malaria as a model, a method which elucidates the profile of antibodies that develop after natural or experimental infection or after vaccination with attenuated organisms, and which identifies immunoreactive antigens of interest for vaccine development or other applications. Expression vectors encoding 250 Plasmodium falciparum (Pf) proteins were generated by PCR/recombination cloning; the proteins were individually expressed with >90% efficiency in Escherichia coli cell-free in vitro transcription and translation reactions, and printed directly without purification onto microarray slides. The protein microarrays were probed with human sera from one of four groups which differed in immune status: sterile immunity or no immunity against experimental challenge following vaccination with radiation-attenuated Pf sporozoites, partial immunity acquired by natural exposure, and no previous exposure to Pf. Overall, 72 highly reactive Pf antigens were identified. Proteomic features associated with immunoreactivity were identified. Importantly, antibody profiles were distinct for each donor group. Information obtained from such analyses will facilitate identifying antigens for vaccine development, dissecting the molecular basis of immunity, monitoring the outcome of whole-organism vaccine trials, and identifying immune correlates of protection.
Doolan
Denise L
DL
Naval Medical Research Center, Silver Spring, MD, USA.
Mu
Yunxiang
Y
Unal
Berkay
B
Sundaresh
Suman
S
Hirst
Siddiqua
S
Valdez
Conrad
C
Randall
Arlo
A
Molina
Douglas
D
Liang
Xiaowu
X
Freilich
Daniel A
DA
Oloo
J Aggrey
JA
Blair
Peter L
PL
Aguiar
Joao C
JC
Baldi
Pierre
P
Davies
D Huw
DH
Felgner
Philip L
PL
D000818:Animals; D000913:Antibodies, Protozoan / Q000096:biosynthesis / Q000276:immunology*; D000953:Antigens, Protozoan / Q000235:genetics / Q000276:immunology; D030562:Databases, Protein; D006801:Humans; D017780:Malaria Vaccines / Q000276:immunology; D016778:Malaria, Falciparum / Q000235:genetics / Q000276:immunology*; D010963:Plasmodium falciparum / Q000235:genetics / Q000276:immunology*; D016133:Polymerase Chain Reaction; D040081:Protein Array Analysis / Q000379:methods*; D040901:Proteomics; D015800:Protozoan Proteins / Q000235:genetics / Q000276:immunology
D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D017780:Malaria Vaccines; D015800:Protozoan Proteins